Journal of global antimicrobial resistance · 2025
Emergence of blaKPC-2-carrying megaplasmid in carbapenem-resistant Pseudomonas aeruginosa ST1076: Genomic and functional insights.
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Chen Jing, Zhu Weidong, Lin Chen, Li Zhen, Ye Haowei, Lou Danfeng
Аннотация
OBJECTIVE: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) poses a critical challenge to clinical treatment due to its multidrug resistance and genomic plasticity. Here, we report the genomic and functional characterization of an ST1076 P. aeruginosa isolate carrying blaKPC-2 on a transferable plasmid.
METHODS: Species identification was performed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and blaKPC-2 was identified by polymerase chain reaction. The characteristics of P. aeruginosa L4884 were analysed by whole-genome sequencing and antimicrobial susceptibility testing. Plasmid characterization was carried out through S1-nuclease pulsed-field gel electrophoresis (S1-PFGE), Southern blotting, transconjugation experiments, and comparative genomic analysis.
RESULTS: The isolate, P. aeruginosa L4884, belonged to sequence type ST1076 and demonstrated resistance to multiple β-lactam antibiotics, including imipenem and meropenem. The blaKPC-2 gene was located on a 394-kb IncP-2 megaplasmid, displaying a conserved genetic structure (IS6100-ISKpn27-blaKPC-2-ISKpn6-klcA). Transconjugation experiments succeeded after repeated efforts, confirming the plasmid's ability to transfer at 27°C.
CONCLUSIONS: This study provides new insights into the genetic basis of carbapenem resistance and underscores the need for enhanced surveillance and containment strategies to prevent the spread of CRPA in clinical settings.
METHODS: Species identification was performed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and blaKPC-2 was identified by polymerase chain reaction. The characteristics of P. aeruginosa L4884 were analysed by whole-genome sequencing and antimicrobial susceptibility testing. Plasmid characterization was carried out through S1-nuclease pulsed-field gel electrophoresis (S1-PFGE), Southern blotting, transconjugation experiments, and comparative genomic analysis.
RESULTS: The isolate, P. aeruginosa L4884, belonged to sequence type ST1076 and demonstrated resistance to multiple β-lactam antibiotics, including imipenem and meropenem. The blaKPC-2 gene was located on a 394-kb IncP-2 megaplasmid, displaying a conserved genetic structure (IS6100-ISKpn27-blaKPC-2-ISKpn6-klcA). Transconjugation experiments succeeded after repeated efforts, confirming the plasmid's ability to transfer at 27°C.
CONCLUSIONS: This study provides new insights into the genetic basis of carbapenem resistance and underscores the need for enhanced surveillance and containment strategies to prevent the spread of CRPA in clinical settings.